RNA干扰在动物体内抗乙型肝炎病毒的效果

Inhibition of hepatitis B virus replication and expression by RNA interference in vivo

目的以乙型肝炎病毒(HBV)C区为靶位,动物实验体内观察RNA干扰抗HBV的效果。方法以流体动力学法建立HBV感染的动物模型,将pcDNA 3．1-HBV和体外细胞实验证明有效的小干扰 RNA(siRNA)尾静脉共注射BALB／c小鼠;用时间分辨免疫荧光分析法检测小鼠血清中乙型肝炎表面抗原(HBsAg)水平,用荧光定量聚合酶链反应法检测血清HBV DNA水平,用逆转录聚合酶链反应法检测 HBV C-mRNA,用免疫组织化学法检测肝组织HBsAg和乙型肝炎核心抗原。结果在小鼠体内,siRNA 能有效抑制HBV的复制和表达,干扰效果至少持续3 d。结论靶向HBV C区的siRNA在动物体内能有效抗HBV。

Objective To evaluate the inhibitory effect of small interfering RNA （siRNA） targeting HBV C gene region on hepatitis B virus （HBV） in vivo. Methods An animal model of HBV infection was developed hydrodynamically, and pcDNA3.1-HBV and siRNA were together injected into the tail vein of the BALB/c mice. HBsAg was analyzed by time-resolved immunofiuorometric assay, HBV DNA was analyzed by fiuorogenic quantitative PCR （FQ-PCR）, HBV C-mRNA was detected by semi-quantitative RT-PCR, and viral specific proteins （HBsAg and HBcAg） in the mice livers were assayed using immunohistochemical staining. Results In the mice, the siRNA effectively inhibited HBV replication and expression compared with the controls. The inhibitive effect of siRNA on HBV lasted at least 3 days. Conclusion These results demonstrate that RNAi can substantially inhibit HBV replication and expression in vivo.